A new nucleic acid–based agent inhibits cytotoxic T lymphocyte–mediated immune disorders

• Published in: Journal of allergy and clinical immunology 2013-09, Vol.132 (3), p.713-722.e11
• Main Authors: Wang, Chuang-Wei, MS, Chung, Wen-Hung, MD, PhD, Cheng, Yi-Fang, PhD, Ying, Nien-Wen, MS, Peck, Konan, PhD, Chen, Yuan-Tsong, MD, PhD, Hung, Shuen-Iu, PhD
• Format: Article
• Language: English
• Subjects: Allergy and Immunology; Alloreactivity; Antigens; Antigens, Differentiation, T-Lymphocyte - genetics; aptamer; Aptamers, Nucleotide - administration & dosage; Biological and medical sciences; Bullous diseases of the skin; CD8; CD8 Antigens - genetics; Cytotoxicity; Deoxyribonucleic acid; Dermatology; DNA; drug hypersensitivity; Flow cytometry; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Graft vs Host Disease - immunology; graft-versus-host disease; granulysin; Humans; Immunopathology; Lymphocytes; Medical sciences; Mortality; nucleic acid–based therapeutics; RNA, Small Interfering - administration & dosage; Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis; Stevens-Johnson syndrome; Stevens-Johnson Syndrome - immunology; Studies; systematic evolution of ligands by exponential enrichment; T cell receptors; T-Lymphocytes, Cytotoxic - immunology; toxic epidermal necrolysis
• ISSN: 0091-6749; 1097-6825
• DOI: 10.1016/j.jaci.2013.04.036

Background Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN), and graft-versus-host disease (GVHD) are distinct immune reactions elicited by drugs or allogeneic antigens; however, they share a pathomechanism with the activation of cytotoxic T lymphocytes (CTLs). CTLs produce cytotoxic proteins, cytokines, chemokines, or immune alarmins, such as granulysin (GNLY), leading to the extensive tissue damage and systemic inflammation seen in patients with SJS/TEN or GVHD. Currently, there is no effective therapeutic agent specific for CTL-mediated immune disorders. Objectives By targeting GNLY+ CTLs, we aimed to develop a nucleic acid–based agent consisting of an anti-CD8 aptamer with GNLY small interfering RNA (siRNA). Methods We performed systematic evolution of ligands using exponential enrichment to select and identify effective anti-CD8 aptamers. We developed an aptamer-siRNA chimera using a “sticky bridge” method by conjugating the aptamer with siRNA. We analyzed the inhibitory effects of the aptamer-siRNA chimera on CTL responses in patients with SJS/TEN or GVHD. Results We identified a novel DNA aptamer (CD8AP17s) targeting CTLs. This aptamer could be specifically internalized into human CTLs. We generated the CD8AP17s aptamer–GNLY siRNA chimera, which showed a greater than 79% inhibitory effect on the production of GNLY by drug/alloantigen-activated T cells. The CD8AP17s aptamer–GNLY siRNA chimera decreased cytotoxicity in in vitro models of both SJS/TEN (elicited by drug-specific antigen) and GVHD (elicited by allogeneic antigens). Conclusions Our results identified a new nucleic acid–based agent (CD8 aptamer–GNLY siRNA chimera) that can significantly inhibit CTL-mediated drug hypersensitivity, such as that seen in patients with SJS/TEN, as well as the alloreactivity seen in patients with GVHD. This study provides a novel therapeutic strategy for CTL-mediated immune disorders.